• Title/Summary/Keyword: dimer formation

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Development of Refolding Process to Obtain Active Recombinant Human Bone Morphogenetic Protein-2 and its Osteogenic Efficacy on Oral Stem Cells

  • Lee, Ji-Hye;Jang, Young-Joo
    • International Journal of Oral Biology
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    • v.42 no.2
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    • pp.71-78
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    • 2017
  • BMP-2 is a well-known TGF-beta related growth factor, having a significant role in bone and cartilage formation. It has been employed to promote bone formation in some clinical trials, and to differentiate mesenchymal stem cells into osteoblasts. However, it is difficult to obtain this protein in its soluble and active form. hBMP-2 is expressed as an inclusion body in the bacterial system. To continuously supply hBMP-2 for research, we optimized the refolding of recombinant hBMP-2 expressed in E. coli, and established an efficient method by using detergent and alkali. Using a heparin column, the recombinant hBMP-2 was purified with the correct refolding. Although combinatorial refolding remarkably enhanced the solubility of the inclusion body, a higher yield of active dimer form of hBMP-2 was obtained from one-step refolding with detergent. The refolded recombinant hBMP-2 induced alkaline phosphatase activity in mouse myoblasts, at $ED_{50}$ of 300-480ng/ml. Furthermore, the expressions of osteogenic markers were upregulated in hPDLSCs and hDPSCs. Therefore, using the process described in this study, the refolded hBMP-2 might be cost-effectively useful for various differentiation experiments in a laboratory.

Different Functional and Structural Characteristics between ApoA-I and ApoA-4 in Lipid-Free and Reconstituted HDL State: ApoA-4 Showed Less Anti-Atherogenic Activity

  • Yoo, Jeong-Ah;Lee, Eun-Young;Park, Ji Yoon;Lee, Seung-Taek;Ham, Sihyun;Cho, Kyung-Hyun
    • Molecules and Cells
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    • v.38 no.6
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    • pp.573-579
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    • 2015
  • Apolipoprotein A-I and A-IV are protein constituents of high-density lipoproteins although their functional difference in lipoprotein metabolism is still unclear. To compare anti-atherogenic properties between apoA-I and apoA-4, we characterized both proteins in lipid-free and lipidbound state. In lipid-free state, apoA4 showed two distinct bands, around 78 and $67{\AA}$ on native gel electrophoresis, while apoA-I showed scattered band pattern less than $71{\AA}$. In reconstituted HDL (rHDL) state, apoA-4 showed three major bands around $101{\AA}$ and $113{\AA}$, while apoA-I-rHDL showed almost single band around $98{\AA}$ size. Lipid-free apoA-I showed 2.9-fold higher phospholipid binding ability than apoA-4. In lipid-free state, $BS_3$-crosslinking revealed that apoA-4 showed less multimerization tendency upto dimer, while apoA-I showed pentamerization. In rHDL state (95:1), apoA-4 was existed as dimer as like as apoA-I. With higher phospholipid content (255:1), five apoA-I and three apoA-4 were required to the bigger rHDL formation. Regardless of particle size, apoA-I-rHDL showed superior LCAT activation ability than apoA-4-rHDL. Uptake of acetylated LDL was inhibited by apoA-I in both lipid-free and lipid-bound state, while apoA-4 inhibited it only lipid-free state. ApoA-4 showed less anti-atherogenic activity with more sensitivity to glycation. In conclusion, apoA-4 showed inferior physiological functions in lipid-bound state, compared with those of apoA-I, to induce more pro-atherosclerotic properties.

Effect of Low Molecular Weight Silk Fibroin on the Inhibition of Tyrosinase Activity

  • Kang, Gyung Don;Lee, Ki Hoon;Shin, Bong Seob;Nahm, Joong Hee;Park, Young Hwan
    • International Journal of Industrial Entomology and Biomaterials
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    • v.9 no.1
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    • pp.29-33
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    • 2004
  • Low molecular weight silk fibroin (LMSF), which was prepared by hydrolysis of silk fibroin using high-temperature and high-pressure method, was found to inhibit the oxidation of L-3,4,-dihydroxyphenylalanine (L-DOPA) catalyzed by mushroom tyrosinase (EC 1.14.18.1). LMSF contained mostly free amino acids such as L-glycine, L-alanine, and L-serine and oligopeptides, mainly glycine-alanine dimer. As a result of analyzing the inhibition kinetics from Lineweaver-Burk plots, L-glycine and glycine-alanine dimer showed noncompetitive behavior while uncompetitive behavior was observed in L-alanine, and L-serine. When weight percent concentration of ${ID_50}$ was compared, L-glycine was most effective on the inhibition and LMSF was also good enough for the inhibition effect of tyrosinase activity. LMSF showed a mixed-type inhibition and the inhibitory mechanism of LMSF might be caused by free amino acids and oligopeptides. As a result of spectroscopic observation with time, initial rate of increase of DOPAchrome decreased remarkably and the time to reach maximum absorbance increased as an increase of the concentration of L-glycine, meaning that L-glycine made itself mainly responsible for the formation of chelate with ${Cu^2+}$ in tyrosinase. However, in case of L-alanine, L-serine, and especially glycine-alanine dimmer, the production of DOPAchrome after an arrival at maximum absorbance decreased, indicating the production of adducts through the reaction with DOPAquinone.

The Continuous Pyrolysis of Waste Polystyrene using Wetted-Wall Type Reactor (Wetted-Wall Column 형 반응기를 이용한 폐 EPS 연속 열분해반응)

  • Han, Myung Sook;Han, Myung Wan;Yoon, Byung Tae;Kim, Seong Bo;Choi, Myoung Jae
    • Korean Chemical Engineering Research
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    • v.45 no.4
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    • pp.396-399
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    • 2007
  • Organic residue and carbonized solid producing from the thermal degradation gave a influence on oil conversion, formation of styrene and side products such as ${\alpha}-methyl$ styrene, ethyl benzene, dimer. Thus, new reaction system using wetted-wall type reactor was proposed and examined on influence of various parameters such as reaction temperature, feeding rate and removal velocity of formed vapor. Optimum condition were obtained from continuous thermal degradation using wetted-wall type reactor and styrene was continuously obtained as the yield up 65%.

In-situ Deposition Rate Measurement System to Improve the Accuracy of the Film Formation Process (성막 공정 정밀도 향상을 위한 실시간 성막 속도 측정 시스템)

  • Somi Park;Seung-Yo Baek;Hyun-Bin Kim;Jonghee Lee;Jae-Hyun Lee
    • Applied Chemistry for Engineering
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    • v.34 no.4
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    • pp.383-387
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    • 2023
  • The quartz crystal microbalance (QCM), commonly used in high vacuum deposition, becomes difficult to use when a thick film is deposited on the quartz, affecting the crystal's inherent vibration. In this study, a non-destructive optical measurement method was developed to measure the film's deposition rate during the in-situ film deposition process. By measuring the scattered laser intensity caused by the dimer in the parylene gas passing through the gas flow path, it was successfully confirmed that the ratio of the dimer in the parylene gas increases as the pyrolysis temperature decreases. Additionally, it was noted that the film's thickness and haze increase as the pyrolysis temperature decreases by confirming the characteristics of the visible parylene films. Through the research results, we aim to utilize the stable in-situ film deposition rate measurement system to control the precise film deposition rate of parylene films.

Analysis of the Dimerization of Human CD99 Using Bimolecular Fluorescence Complementation Technique

  • Lee, Mi-Kyung;Kim, Hyun-Soo;Kim, Seung-Seok;Cho, Myung-Hwan;Lee, Im-Soon
    • Journal of Microbiology and Biotechnology
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    • v.18 no.3
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    • pp.472-476
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    • 2008
  • Two isoforms of human CD99 have been identified, but only heterotypic interaction between the isomers was recently demonstrated. In this study, we performed bimolecular fluorescence complementation analysis to further characterize the interaction in vivo. Upon transiently transfecting plasmids expressing either of the two isoforms fused with yellow fluorescent protein (YFP) fragments, all the YFP-tagged CD99 molecules were properly localized on cell surfaces, and formed fluorescent dimers. Interestingly, however, unlike the previous report, the homodimers formed as efficiently as the heterodimer via their extracellular domains, implying its distinct regulatory role through modulating the complex profile.

Physical Properties of the Langmuir-Blodgett Films Layered with Octadecylviologen-TCNQ (Octadecylviologen-TCNQ LB 막의 물리적 특성)

  • 신동명;박제상;강도열
    • Proceedings of the Korean Institute of Electrical and Electronic Material Engineers Conference
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    • 1995.05a
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    • pp.77-80
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    • 1995
  • Enhancing the electrical conductivity of the ultrathin organic films using Langmuir-Blodget technique is important step for the developement of molecular electronic device. The Octadecylviologen-TCNQ was synthesized with Octadecylviologen-Bromide an Lithium TCNQ Sine Octadecylviologen-TCNQ has two TCNQ snion radicals, the conductivity of LB film is expected to increase. The $\pi$-A isotherm showed that the limiting area was 150${\AA}$$\^$2/ molecule and the silid-like transition surface pressure was 25 mN/m. The electronic transition of the TNCNQ anion radical was observed at 400 nm. Intermolecular charge transfer absorption was observed at 600nm and 850~1050 nm which ay resulted from the TCNQ anion radical dimer formation. The electrical conductivity of the viologen -TCNQ LB film was 10$\^$-6/cm This values was 100 times higher than that of the quinolinium-TCNQ and pyridinium-TCNQ LB films.

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Catalytic Activity of Supported Rhodium(I) Complex for the Carbonylation of Nitrobenzene: Mechanism for Carbamate Formation

  • Kim, Jin-Hyung;Kim, Dae-Won;Cheong, Min-Serk;Kim, Hoon-Sik;Mukherjee, Deb Kumar
    • Bulletin of the Korean Chemical Society
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    • v.31 no.6
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    • pp.1621-1627
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    • 2010
  • The investigation of the catalytic activity of supported rhodium(I) complex [Rh(P-S)$(CO)_2$] (P-S; polymer anchored salicylic acid) toward the reductive carbonylation of nitrobenzene in DMF medium has been reported. Use of basic cocatalysts in the reaction medium enhanced the percentage of more useful phenyl carbamates. Spectroscopic studies indicate that the reaction proceeds through a dimer species [Rh(HS)(CO)(C(O)$OCH_3$)(${\mu}-OCH_3)]_2$ and phenyl isocyanate is formed as an intermediate. A plausible reaction mechanism based on the identification of reactive intermediates from the soluble rhodium variety has been proposed for the carbonylation process.

Anion Photoelectron Spectroscopy and Theoretical Calculation of the Hetero-dimers of Polycyclic Aromatic Hydrocarbons

  • Kim, Namdoo;Lee, Sang Hak
    • Bulletin of the Korean Chemical Society
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    • v.34 no.5
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    • pp.1441-1444
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    • 2013
  • Hetero-dimer anions of naphthalene (Np), anthracene (An), phenanthrene (Ph) and pyrene (Py) were investigated using the time-of-flight mass spectrometer (TOF-MS), anion photoelectron spectroscopy (PES) and theoretical calculation. There are two possible geometries with their electron affinity (EA) difference: parallel displaced (PD) and T-shaped. Dispersion force plays a key role in PD structure with the formation of a new anionic core while ${\pi}$-hydrogen interaction plays a key role in T-shaped structure with the monomer anionic core. The optimized structures and charge distributions can simply be explained by the relative difference of EA.

Hydroxy-Substituted Polyenaminonitrile as a Soluble Precursor for Rigid-Rod Polybenzoxazole

  • Kim, Ji Heung;Lee, Jae Gwan
    • Bulletin of the Korean Chemical Society
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    • v.22 no.9
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    • pp.999-1004
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    • 2001
  • (1-Chloro-2,2-dicyanovinyl)benzene or 1,4-bis(1-chloro-2,2-dicyanovinyl)benzene was reacted with 2-amino-phenol to give the model compound, hydroxy enaminonitrile, which was found to undergo thermal cyclization reaction to form the corresponding benzoxazole. This intramolecular cyclization reaction is expected to occur through nucleophilic attack to electropositive enamine carbon by ortho-hydroxy group on the phenyl ring, which is accompanied by the release of neutral malononitrile through rearrangement. From each bifunctional monomer, o-hydroxy substituted polyenaminonitrile was prepared and characterized as a new precursor polymer for well-known aromatic polybenzoxazole. Also the unusual macrocyclic dimer formation from the 1,4-bis(1-chloro-2,2-dicyanovinyl)benzene and 2,2-bis(3-amino-4-hydroxyphenyl)hexafluoropropane polymerization reaction system was discussed. The thermal cyclization reactions and the properties of polymers were investigated using FT-IR and thermal analysis (DSC & TGA).